Purification and properties of extracellular carboxyl proteases of acid-tolerant bacteria, isolated from compost

• Published in: Bioscience, biotechnology, and biochemistry Vol. 63; no. 11; pp. 1927 - 1933
• Main Authors: Takehana, T. (Hokkaido Univ., Sapporo (Japan)), Inoue, S, Takei, R, Ito, H, Matsui, H, Honma, M
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01-11-1999; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: ACIDE; ACIDOS; ACIDS; Amino Acid Sequence; ANALISIS BIOLOGICO; ANALYSE BIOLOGIQUE; BACTERIA GRAM NEGATIVA; BACTERIE GRAM NEGATIF; BIOLOGICAL ANALYSIS; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Burkholderia; carboxyl protease; Carboxypeptidases - chemistry; Carboxypeptidases - isolation & purification; Carboxypeptidases - metabolism; CELLS; CELLULE; CELULAS; Chromatography, Gel; Chromatography, Ion Exchange; COMPOST; COMPOSTS; Fundamental and applied biological sciences. Psychology; gel filtration; GRAM NEGATIVE BACTERIA; Gram-Negative Aerobic Rods and Cocci - enzymology; Gram-Negative Aerobic Rods and Cocci - growth & development; Hydrogen-Ion Concentration; Kinetics; Miscellaneous; Mission oriented research; Molecular Sequence Data; Molecular Weight; pepstatin; Peptide Fragments - chemistry; PROTEASAS; PROTEASE; PROTEASES; Proteobacteria; Proteobacteria - enzymology; Proteobacteria - growth & development; PSEUDOMONAS; PURIFICACION; PURIFICATION; RESISTANCE AUX PRODUITS CHIMIQUES; RESISTANCE TO CHEMICALS; RESISTENCIA A PRODUCTOS QUIMICOS; Sequence Alignment; Sequence Homology, Amino Acid; Soil Microbiology; Stenotrophomonas; Pseudomonadales; Compost; Purification; Enzyme; Stenotrophomonas; Enzyme inhibitor; Identification; Extracellular; Peptidases; Xanthomonapepsin; Enzymatic activity; Bacteria; Hydrolases; Pseudomonadaceae; Isolation; Aspartic endopeptidases; Burkholderia; Physicochemical properties
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.63.1927

Four strains of acid-tolerant and protein-using bacteria were isolated from compost. Two of them, Gram-negative strains MB8 and MB11, were identified as a new genus close to Stenotrophomonas species MB8 and Burkholderia species MB11, respectively. Both bacteria produced extracellular carboxyl proteases (CP) in acid-casein-starch medium. The enzymes, termed CP MB8 and CP MB11, purified through ion exchange and gel filtration chromatographies had molecular weights of 61,000 (CP MB8) and 36,000 (CP MB11) as estimated by SDS-PAGE, and showed optimum activities with hemoglobin as a substrate at pH 3.5 (CP MB8) and pH 3.7 (CP MB11) at 55 degrees C. Both of the enzymes were not inhibited by pepstatia, DAN, or EPNP. These results suggest that both enzymes are members of the pepstatin-insensitive carboxyl proteinase family (EC 3.4.23.33), except for a larger molecular weight of the CP MB8 enzyme